LOCUS       sequence-131174-        7778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Netrin G1, isoform e 
            protein (truncated immediately upstream of propeptide cleavage and 
            GPI-attachement site), C-terminally fused to alkaline phosphatase + 
            6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7778)
  AUTHORS   Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray 
            TA, Kay JN, Wojtowicz WM
  TITLE     An extracellular biochemical screen reveals that FLRTs and Unc5s 
            mediate neuronal subtype recognition in the retina.
  JOURNAL   Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149.
  PUBMED    26633812
REFERENCE   2  (bases 1 to 7778)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Ntng1.e-AP-His
FEATURES             Location/Qualifiers
     source          1..7778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     609..633
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1573
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             4358..4375
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4401..4522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4438..4457)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4492..4511
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4560..4577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4731..5319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4811..4830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5490..6281,6282..6350))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6113..6132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6351..6455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6523..6541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6579..6601)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6603..6623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6608..6938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6789..6924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6851..6870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7075..7094
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7161..7616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7298..7319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7510..7529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7742..7764
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7756..7773
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7757..7773
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc
      781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc
      841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
      901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
      961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta
     1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc
     1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt
     1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc
     1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac
     1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt
     1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta
     1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg
     1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg
     1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg
     1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tgtatttgtc
     1621 aagattcctg tcgatccatg ccctgtgggt gacagtgtcc tctgtgatgc agccctacct
     1681 tttcgtgtgg ggacattatg atgtatgtaa gagcctgatt tacacagaag aaggcaaagt
     1741 ttgggattac acagcctgcc agccggaatc cacggacatg accaagtatc tgaaagtgaa
     1801 actggaccct ccggatatta cctgtggaga ccctccagag tccttctgtg caatgggcaa
     1861 cccttacatg tgcaataatg agtgtgatgc gagtacccct gaactggcac accctcctga
     1921 gctgatgttt gattttgaag gaagacatcc ctccacattt tggcagtctg ctacttggaa
     1981 ggagtacccc aaacctctcc aggttaacat cactctgtct tggagcaaaa ccattgaact
     2041 cacagacaac atagttatta cctttgaatc ggggcgtcca gaccaaatga tcctagagaa
     2101 atctctcgac tacggacgaa catggcagcc ctatcagtat tatgccacag actgcctcca
     2161 tgcattccac atggacccga aatccgtgaa ggatttatct cagcacacgg tcttggaaat
     2221 catttgcacg gaagagtact ccactgggta ctccacgaat agcaaaataa tccacttcga
     2281 gatcaaagac aggtttgcgt ttttcgctgg acctcggcta cgaaatatgg cttccctcta
     2341 tggacagctg gatacaacca agaaactcag agatttcttc actgtcacag acctgaggat
     2401 caggctgttg agacccgccg ttggggaaat atttgtagat gaactacatt tggcacgtta
     2461 cttttatgcg atctcagaca taaaggtgcg aggaaggtgc aagtgcaacc tgcatgccac
     2521 ttcgtgtttg tatgacaaca gcaaactgac atgtgaatgt gagcacaaca ctacaggtcc
     2581 cgactgtggg aaatgcaaga agaactacca gggccgacct tggagccccg gctcatacct
     2641 ccccatcccc aaaggcaccg caaacacctg tatccccagc atttccagta tcggtactcc
     2701 tccaaagttt aataggatat ggccgaatat ttcttccctt gaggtttcta acccaaaaca
     2761 agctaatgtc tgcgacaatg agctcctgca ctgccagaat ggagggacct gccagaacaa
     2821 tgtgcgctgc gcgtgcccag acgcctacac cggcatcctc tgtgagaagc taactagtgg
     2881 atcttccgga atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     2941 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     3001 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     3061 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     3121 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     3181 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     3241 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     3301 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     3361 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     3421 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     3481 cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     3541 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     3601 gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc
     3661 cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga
     3721 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     3781 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     3841 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     3901 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     3961 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     4021 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     4081 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     4141 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     4201 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     4261 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     4321 gccccccgcc ggcaccaccg acgccgcgca cccgggtcat catcaccatc accattaatc
     4381 tagagatctt attaaagcag aacttgttta ttgcagctta taatggttac aaataaagca
     4441 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4501 ccaaactcat caatgtatct tatcatgtct ggtcgactct agctagactc ttccgcttcc
     4561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5161 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5641 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5881 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5941 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6001 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6061 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6121 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6181 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6241 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6301 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6481 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6541 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6601 gaggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta
     6661 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6781 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6841 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     6901 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     6961 ccattttcgt cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7021 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7081 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     7141 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     7201 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7261 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7321 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7381 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7441 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7501 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7561 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     7621 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7681 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7741 tcccagtcac gacgttgtaa aacgacggcc agtgaatt
//
